It is desirable to lyse cells grown as production hosts containing a protein or other molecule of interest to recover any desired intracellularly produced product. Conventional ways to kill and lyse such cells include the use of heat (U.S. Pat. No. 4,601,986 to Wegner, et al.), osmotic pressure (U.S. Pat. No. 4,299,858 to Aubert, et al), enzymes which break down the cell walls or membranes (U.S. Pat. No. 3,816,260 to Sugiyama, U.S. Pat. No. 3,890,198 to Kobayashi, et al. and U.S. Pat. No. 3,917,510 to Kitamura, et al.) and mechanical disruption of the cell wall by, for example, high pressure homogenization. The disclosures of the above patents are incorporated herein by reference.
Also, detergents have been utilized to lyse the cell wall. For example, yeast protein extraction reagent (Y-PER®), sold by Pierce Chemical Company, contains a detergent to provide a gentle means of cell lysis that is not detrimental to the protein of interest. However, Y-PER® is intended to be used as a laboratory bench reagent, not as a reagent useful for the large scale production of proteins, and is costly. For these reasons, Y-PER® has not gained acceptance as a useful reagent for the large scale production of recombinant protein from host cells.
There is a need in the art for a process that can be used to easily cause the release of proteins from host cells without harming the desired protein and with a minimum of process steps. The method of cell lysis should not directly or indirectly lead to denaturation of the desired product and the method should be consistent with subsequent processing requirements and with large scale production.